Departments of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Invest Ophthalmol Vis Sci. 2010 Apr;51(4):1823-31. doi: 10.1167/iovs.09-4657. Epub 2009 Nov 20.
Purpose. MicroRNAs (miRNAs) are short, noncoding transcripts that negatively regulate gene expression. They are implicated in diverse cellular processes. The purpose of this study was to obtain a global expression profile of miRNAs in the developing retina and identify differences in miRNA expression between adult rod and cone photoreceptors. Methods. Locked nucleic acid (LNA) microarrays were used to investigate the miRNA transcriptome of the developing mouse retina and brain. Real-time PCR was used to validate the array findings. Laser capture microdissection was used to determine the miRNA spatial pattern of expression. Results. One hundred thirty-eight miRNAs were expressed at at least one of the investigated time points. Several miRNAs showed significant changes in expression between embryonic day 15 and adult age in both retina and brain. Cluster analysis identified subgroups of miRNAs showing defined expression profiles. Globally, correlation of expression was higher, with increasing sequence similarity of the mature miRNAs. The miRNAs with identical seed sequences exhibited highly correlated expression profiles. The co-expression of selected host gene and intronic miRNA pairs was confirmed in adult retina. In some cases, expression profiles of miRNAs showed weak correlation with those of their host transcripts, suggesting posttranscriptional regulation of miRNAs during development. In addition, the miRNA transcriptome of rod- and cone-dominant retinas showed only minor differences, and no miRNAs specific for either cell-type were identified. Conclusions. Global expression profiling revealed dozens of miRNAs with significant expression changes in the developing retina. Precise patterns of expression of miRNAs suggest their specific roles in development.
MicroRNAs(miRNAs)是一种短的非编码转录物,能够负向调控基因表达。它们参与多种细胞过程。本研究的目的是获得发育中视网膜中 miRNAs 的全局表达谱,并鉴定成年视杆和视锥光感受器中 miRNA 表达的差异。
采用锁核酸(LNA)微阵列来研究发育中的小鼠视网膜和大脑中的 miRNA 转录组。实时 PCR 用于验证芯片结果。激光捕获显微切割用于确定 miRNA 空间表达模式。
在至少一个研究时间点,有 138 个 miRNAs 表达。在视网膜和大脑中,有几个 miRNAs 在胚胎期 15 天到成年期之间的表达有显著变化。聚类分析确定了具有特定表达谱的 miRNAs 亚群。总的来说,随着成熟 miRNAs 序列相似性的增加,表达的相关性更高。具有相同种子序列的 miRNAs 表现出高度相关的表达谱。在成年视网膜中,选定的宿主基因和内含子 miRNA 对的共表达得到了证实。在某些情况下,miRNAs 的表达谱与宿主转录物的表达谱相关性较弱,表明 miRNA 在发育过程中受到转录后调控。此外,视杆和视锥优势视网膜的 miRNA 转录组仅显示出微小差异,并且没有鉴定出特定于任何一种细胞类型的 miRNAs。
全局表达谱分析揭示了发育中视网膜中数十个具有显著表达变化的 miRNAs。miRNAs 的精确表达模式表明它们在发育过程中的特定作用。